Based on the strategy of metabolomics combined with bioinformatics, this study analyzed the potential allergens and mechanism of pseudo-allergic reactions (PARs) induced by the combined use of Reduning injection and penicillin G injection. All animal experiments and welfare are in accordance with the requirements of the First Affiliated Experimental Animal Ethics and Animal Welfare Committee of Henan University of Chinese Medicine (approval number: YFYDW2020002). Based on UPLC-Q-TOF/MS technology combined with UNIFI software, a total of 21 compounds were identified in Reduning and penicillin G mixed injection. Based on molecular docking technology, 10 potential allergens with strong binding activity to MrgprX2 agonist sites were further screened. Metabolomics analysis using UPLC-Q-TOF/MS technology revealed that 34 differential metabolites such as arachidonic acid, phosphatidylcholine, phosphatidylserine, prostaglandins, and leukotrienes were endogenous differential metabolites of PARs caused by combined use of Reduning injection and penicillin G injection. Through the analysis of the "potential allergen-target-endogenous differential metabolite" interaction network, the chlorogenic acids (such as chlorogenic acid, neochlorogenic acid, cryptochlorogenic acid, and isochlorogenic acid A) and β-lactam allergens in the combination of the two may be mainly regulated by PLD1, PLA2G12A and CYP1A1. The three upstream signal target proteins mainly activate the arachidonic acid metabolic pathway, promote the degranulation of mast cells, release downstream endogenous inflammatory mediators, and induce PARs.

Squalene epoxidase (SQLE) is a potential target for the treatment of liver cancer. Bioinformatics analysis indicated that the high expression of SQLE was closely related to the clinical stage and poor prognosis of patients with liver cancer. However, the existing inhibitors against SQLE 195 tyrosine residue (Y195) cannot be used clinically due to severe side effects. In this study, 35 small-molecule compounds targeting SQLE 335 tyrosine residue (Y335) were selected by computer virtual screening. Combined with MTT assay, 3 candidate compounds (19^#, 31^# and 35^#) with significant inhibitory effects on the proliferation of Huh7 cell line were obtained. Further studies showed that these 3 compounds could inhibit the migration of Huh7 cells, reduce the contents of total and free cholesterol, up-regulate the expression of tumor suppressor gene PTEN, and down-regulate the expression of PI3K and AKT proteins. The results showed that the novel inhibitors 19^#, 31^# and 35^# targeting SQLE Y335 could reduce cholesterol content, inhibit the proliferation and migration of Huh7, thus playing an anti-liver cancer role.

Objective To investigate the effect of total alkaloids of Rhizoma Corydalis(TAC)on the expression of silencing information regulator 1(Sirt1)/tumor suppressor P53 protein signaling pathway-related proteins in the hippocampus of rats with chronic cerebral ischemia(CCH),and to explore its mechanism.Methods The rats were randomly divided into Sham operation group,model group,TAC high-dose group(14 mg/kg)and TAC low-dose group(7 mg/kg),with 6 rats in each group.A modified bilateral common carotid artery permanent occlusion method(BCCAO)was used to establish a rat model of CCH,and only bilateral common carotid arteries were separated in the Sham group.After the modeling was completed,each group was given the corresponding drug or isotonic saline by gavage,once a day,and the treatment lasted for 14 days.Hematoxycin-eosin staining was used to observe the pathological changes in the hippocampus of rats,in situ terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphate-biotin nick end labeling assay(TUNEL)was used to detect neuronal apoptosis,and Western blotting and immunohistochemistry were used to detect expression of Sirt1,P53,P53 positive apoptosis regulator(PUMA),B-cell lymphocytoma-2(Bcl-2)protein,Bcl-2-related X protein(BAX),respectively in the hippocampus of rats.Results(1)There were significant differences in the number of apoptotic cells and apoptosis rate among the four groups(F-values were 71.417 and 76.835,respectively,both P＜0.01).There were statistically significant differences in the mean integral optical density values of Sirt1,P53,PUMA,BAX and Bcl-2 protein positive expression areas among the four groups(F-values were 1 178.390,42.465,867.413,110.656 and 131.801,all P＜0.01).There were significant differences in the relative expression levels of Sirt1,P53,PUMA,BAX and Bcl-2 among the four groups(F-values were 9.497,11.863,58.552,186.855 and 12.466,all P＜0.01).(2)Compared with the Sham operation group,the neuronal arrangement of brain tissue in the hippocampus of the model group was disordered,the nuclear consolidation increased,and the glial cells and inflammatory cells increased significantly,and the number and apoptosis rate of neurons in the hippocampus of the model group increased significantly(respectively[10.8±1.5]cells vs.[2.0±0.9]cells and[35.5±4.5]％vs.[6.2±2.6]％;both P＜0.05),and the average integral optical density values of the positive expression areas of Sirt1 and Bcl-2 proteins decreased significantly(84.6±6.6 vs.244.6±4.9,138.5±6.7 vs.210.9±10.0;both P＜0.05),the average integral optical density values of P53,PUMA and BAX proteins were significantly increased(156.8±11.6 vs.93.5±11.6,151.3±3.3 vs.38.0±4.0,87.0±5.0 vs.38.4±5.5;all P＜0.05),the relative expression levels of Sirt1 and Bcl-2 proteins were significantly decreased(0.51±0.07 vs.0.74±0.07,0.36±0.03 vs.0.53±0.05;both P＜0.05),and the relative expression levels of P53,PUMA and BAX proteins were significantly increased(0.37±0.06 vs.0.21±0.02,0.62±0.06 vs.0.23±0.02,1.08±0.06 vs.0.45±0.03;all P＜0.05).(3)Compared with the model group,the hippocampal tissue structure of the high-dose and low-dose TAC groups was relatively compact and uniform,the neurons were neatly arranged,and the cell structure was relatively clear and complete,while the number of neuronal apoptotic cells and the apoptosis rate decreased significantly(respectively[3.8±0.7]cells vs.[6.2±1.2]cells,[12.4±2.8]％vs.[20.2±3.9]％;both P＜0.05),and the average integrated optical density values of the positive expression areas of Sirt1 and Bcl-2 proteins(the high-dose and low-dose TAC groups:Sirt1 150.0±4.8,131.3±1.3,and Bcl-2 207.1±7.4,169.5±3.9,respectively)were significantly increased(both P＜0.05),the average integral optical density values of P53,PUMA and BAX proteins were significantly decreased(the high-dose and low-dose TAC groups:P53 105.9±8.8,115.5±9.0,and PUMA56.8±5.1,74.4±3.9,and BAX40.5±5.6,48.4±5.0,respectively,all P＜0.05),the relative expression levels of Bcl-2 protein(the high-dose and low-dose TAC groups:0.53±0.05,0.47±0.02,respectively)were significantly increased(P＜0.05),the relative expression levels of P53(the high-dose and low-dose TAC groups:0.21±0.02,0.24±0.04,respectively),PUMA(the high-dose and low-dose TAC groups:0.36±0.02,0.28±0.04,respectively)and BAX proteins(the high-dose and low-dose TAC groups:0.52±0.02,0.54±0.03,respectively)were significantly decreased(all P＜0.05),the relative expression level of Sirt1 protein in the TAC high-dose group was significantly decreased(0.71±0.05,P＜0.05),and the relative expression level of Sirt1 protein in the TAC low-dose group was not statistically significant(0.52±0.08,P＞0.05).Conclusion TAC can alleviate neuronal damage and reduce the apoptosis rate of neurons in the hippocampus of CCH rats,and the mechanism may be related to the activation of Sirt1/P53 pathway,inhibition of P53 protein activity,and thus the expression level of apoptosis-related proteins in the downstream of TAC.

Transparency Ecosystem for Research and Journals in Medicine (TERM) working group summarized the essential recommendations that should be considered to review and publish a high-quality guideline. These recommendations from editors and reviewers included 10 components of essential requirements: systematic review of existing relevant guidelines, guideline registration, guideline protocol, stakeholders, conflicts of interest, clinical questions, systematic reviews, recommendation consensus, guideline reporting and external review. TERM working group abbreviates them as PAGE (essential requirements for Publishing clinical prActice GuidelinEs), and recommends guideline authors, editors, and peer reviewers to use them for high-quality guidelines.
Humans ; Practice Guidelines as Topic

OBJECTIVE: KRAS gene is one of the most common mutations of proto-oncogenes in human tumors, G12V is one of the most common mutation types for KRAS. It's challenging to chemically acquire the targeted drug for this mutation. Recent studies reported that this mutation peptides can form a neoepitope for T cell recognition. Our study aims to clone the T cell receptor (TCR) which specifically recognizes the neoepitope for KRAS G12V mutation and constructs TCR engineered T cells (TCR-T), and to investigate if TCR-Ts have strong antitumor response ability. METHODS: In this study, tumor infiltrating lymphocytes were obtained from one colorectal cancer patient carrying KRAS G12V mutation. Tumor-reactive TCR was obtained by single-cell RT-5' rapid-amplification of cDNA ends PCR analysis and introduced into peripheral blood lymphocytes to generate TCR-Ts. RESULTS: We obtained a high-affinity TCR sequence that specifically recognized the HLA-A^*11:01-restricted KRAS G12V_8-16 epitope: KVA11-01. KVA11-01 TCR-T could significantly kill various tumor cells such as PANC-1, SW480 and HeLa (overexpressing HLA-A^*11:01 and KRAS G12V), and secreting high levels of interferon-γ (IFN-γ). Non-specific killing experiments suggested KVA11-01 specifically recognized tumor cells expressing both mutant KRAS G12V and HLA-A^*11:01. In vivo assay, tumor inhibition experiments demonstrated that infusion of approximately 1E7 KVA11-01 TCR-T could significantly inhibit the growth of subcuta-neously transplanted tumors of PANC-1 and HeLa (overexpressing HLA-A^*11:01 and KRAS G12V) cells in nude mice. No destruction of the morphologies of the liver, spleen and brain were observed. We also found that KVA11-01 TCR-T could significantly infiltrate into tumor tissue and had a better homing ability. CONCLUSION KVA11-01 TCR-T cells can effectively target a variety of malignant tumor cells carrying KRAS G12V mutation through in vitro and in vivo assay. KVA11-01 TCR-T cells have excellent biological activity, high specificity of target antigen and homing ability into solid tumor tissue. KVA11-01 TCR-T is expected to be an effective treatment for patients with KRAS G12V mutant solid malignancies.
Animals ; DNA, Complementary ; Epitopes ; HLA-A Antigens ; Humans ; Interferon-gamma ; Mice ; Mice, Nude ; Mutation ; Neoplasms ; Proto-Oncogene Proteins p21(ras)/genetics* ; Receptors, Antigen, T-Cell/genetics*

Objective: To explore the effect of perioperative chemotherapy on the prognosis of gastric cancer patients under real-world condition. Methods: A retrospective cohort study was carried out. Real world data of gastric cancer patients receiving perioperative chemotherapy and surgery + adjuvant chemotherapy in 33 domestic hospitals from January 1, 2014 to January 31, 2016 were collected. Inclusion criteria: (1) gastric adenocarcinoma was confirmed by histopathology, and clinical stage was cT2-4aN0-3M0 (AJCC 8th edition); (2) D2 radical gastric cancer surgery was performed; (3) at least one cycle of neoadjuvant chemotherapy (NAC) was completed; (4) at least 4 cycles of adjuvant chemotherapy (AC) [SOX (S-1+oxaliplatin) or CapeOX (capecitabine + oxaliplatin)] were completed. Exclusion criteria: (1) complicated with other malignant tumors; (2) radiotherapy received; (3) patients with incomplete data. The enrolled patients who received neoadjuvant chemotherapy and adjuvant chemotherapy were included in the perioperative chemotherapy group, and those who received only postoperative adjuvant chemotherapy were included in the surgery + adjuvant chemotherapy group. Propensity score matching (PSM) method was used to control selection bias. The primary outcome were overall survival (OS) and progression-free survival (PFS) after PSM. OS was defined as the time from the first neoadjuvant chemotherapy (operation + adjuvant chemotherapy group: from the date of operation) to the last effective follow-up or death. PFS was defined as the time from the first neoadjuvant chemotherapy (operation + adjuvant chemotherapy group: from the date of operation) to the first imaging diagnosis of tumor progression or death. The Kaplan-Meier method was used to estimate the survival rate, and the Cox proportional hazards model was used to evaluate the independent effect of perioperative chemo therapy on OS and PFS. Results: 2 045 cases were included, including 1 293 cases in the surgery+adjuvant chemotherapy group and 752 cases in the perioperative chemotherapy group. After PSM, 492 pairs were included in the analysis. There were no statistically significant differences in gender, age, body mass index, tumor stage before treatment, and tumor location between the two groups (all P>0.05). Compared with the surgery + adjuvant chemotherapy group, patients in the perioperative chemotherapy group had higher proportion of total gastrectomy (χ(2)=40.526, P<0.001), smaller maximum tumor diameter (t=3.969, P<0.001), less number of metastatic lymph nodes (t=1.343, P<0.001), lower ratio of vessel invasion (χ(2)=11.897, P=0.001) and nerve invasion (χ(2)=12.338, P<0.001). In the perioperative chemotherapy group and surgery + adjuvant chemotherapy group, 24 cases (4.9%) and 17 cases (3.4%) developed postoperative complications, respectively, and no significant difference was found between two groups (χ(2)=0.815, P=0.367). The median OS of the perioperative chemotherapy group was longer than that of the surgery + adjuvant chemotherapy group (65 months vs. 45 months, HR: 0.74, 95% CI: 0.62-0.89, P=0.001); the median PFS of the perioperative chemotherapy group was also longer than that of the surgery+adjuvant chemotherapy group (56 months vs. 36 months, HR=0.72, 95% CI:0.61-0.85, P<0.001). The forest plot results of subgroup analysis showed that both men and women could benefit from perioperative chemotherapy (all P<0.05); patients over 45 years of age (P<0.05) and with normal body mass (P<0.01) could benefit significantly; patients with cTNM stage II and III presented a trend of benefit or could benefit significantly (P<0.05); patients with signet ring cell carcinoma benefited little (P>0.05); tumors in the gastric body and gastric antrum benefited more significantly (P<0.05). Conclusion: Perioperative chemotherapy can improve the prognosis of gastric cancer patients.
Chemotherapy, Adjuvant ; Female ; Gastrectomy ; Humans ; Male ; Neoadjuvant Therapy ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Stomach Neoplasms/surgery*

OBJECTIVETo identify the factors that affect the safety and efficacy of peroral endoscopic myotomy (POEM) for treatment of achalasia.

METHODSData of consecutive patients undergoing POEM for confirmed achalasia between December, 2010 and December, 2015 were collected, including the procedure time, approach of tunnel entry incision, approach of myotomy, complications and follow-up data.

RESULTSAmong the total of 439 patients enrolled, the overall complication rate was 28.7% (126/439). Treatment success (Eckardt score≤3) was achieved in 94.5% of 364 patients followed up for a median of 6 months (1-48 months), and the mean score was reduced significantly from 6.7∓1.5 before treatment to 1.2∓1.1 after the treatment (P<0.05). Logistic regression revealed that the year when POEM was performed and the approach of entry incision were two significant factors contributing to complications: with the year 2015 as the reference, the odds ratio (OR) was 9.454 (95% CI: 2.499-35.76) for the years before 2011, 2.177 (95% CI: 0.794-5.974) for 2012, 3.975 (95% CI: 1.904-8.298) for 2013, and 1.079 (95% CI: 0.601-1.940) for 2014; with the longitudinal entry incision as the reference, the OR was 0.369 (95% CI: 0.165-0.824) for inverted T entry incision and 0.456 (95% CI: 0.242-0.859) for transverse entry incision. The approach of myotomy was the significantly associated with symptomatic relapse: with full-thickness myotomy combined with indwelling an anti-reflux belt as the reference, the OR was 0.363 (95% CI: 0.059-2.250) for gradual full-thickness myotomy, 2.137 (95% CI: 0.440-10.378) for circular muscle myotomy, and 4.385 (95% CI: 0.820-23.438) for circular muscle myotomy in combination with balloon shaping; the recurrence rate was 0 with a full-thickness myotomy.

CONCLUSIONThe complication rates of POEM appears to decrease over time, and an inverted T entry incision is the best choice for controlling the complications. Gradual full-thickness myotomy is an excellent approach for treatment of achalasia in terms of the relapse rate, procedure time and the incidence of reflux esophagitis.

Endoscopy ; Esophageal Achalasia ; surgery ; Esophagitis, Peptic ; surgery ; Gastroesophageal Reflux ; Humans ; Muscles ; surgery ; Recurrence ; Treatment Outcome

OBJECTIVETo investigate the effects of naringin on the proliferation, differention and maturaion of rat calvarial osteoblasts (ROB).

METHODSegregated neonatal SD rat skull, enzyme digestion to obtain ROB. The culture medium was replaced every three days. Serial subcultivation proceeded when cells covered with 80% culture dish. Naringin supplemented into the culture at 1 x 10(-4), 1 x 10(-5), 1 x 10(-6), 1 x 10(-7) mol x L(-1) respectively. MTT method was adopted in proliferation analysis and the activity of ALP was examined after induced 9 days. Search the best concentration and supplemented into the medium, then the osteogenic differentiation markers including the secretion amount of osteocalcin, osteopontin and bone morphogenetic protein-2 were compared between the naringin-supplemented group and the control. Total RNA was isolated and the mRNA level of bFGF, IGF-1, Runx-2, Osterix, ERa and ERbeta was investigated by Real time RT-PCR. Total protein also was isolated and the expression ERa, ERbeta and collagen I was examined by Western blot. After the addition of ICI 182.780, an inhibitor of the estrogen signal pathway, these index also was examined and the changes were compared.

RESULTThe ROB proliferation was motivated by naringin dose-dependently. And it evidently leads to osteogenic process and maturation. 1 x 10(-5) mol x L(-1) is the best concentration. Naringin improved the secretion of osteocalcin, osteopontin, bone morphogenetic protein-2 and collagen I significantly. Besides, it can also enhanced the mRNA level of bFGF, IGF-1, Runx-2, Osterix, ERalpha and ERbeta. While all these effects can be restrained by ICI 182.780.

CONCLUSIONThe naringin with final concentration of 1 x 10(-5) mol x L(-1) enhances the osteogenic differentiation and maturation of ROB significantly, while the promoting effects vanished after the addition of ICI 182.780. These results suggesting that naringin is one of the phytoestrogens and have the activity of bone formation may via estrogen signal pathway, it can be developed into a new drug for osteoporosis therapy.

Alkaline Phosphatase ; genetics ; metabolism ; Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Flavanones ; pharmacology ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteocalcin ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Skull ; cytology ; drug effects ; metabolism

OBJECTIVETo explore the influence of compound whole grain complex antioxidant chain on oxidative stress to the hyperlipidemia population.

METHODSFrom March 2008 to March 2009, 418 hyperlipemia residents (45 to 75 years of age) of Han group were screened from 3 main districts in Nanjing, according to the community, blood lipids and oxidative indicators, stratified into intervention and control group by quasi-experimental design. The intervention group (212 individuals) were provided with compound whole grain and health education while only health education was provided for the control group (206 individuals). Body mass index (BMI), waist-to-hip ratio (WHR), total cholesterol (TC), triglyceride (TG) and high density lipoprotein cholesterol (HDL-C) and oxidative indicators (including T-AOC, MDA, SOD, GSH-Px) were measured before and after the one-year intervention period. Analyses of the correlation between posture, biochemical markers and oxidative stress indicators before and after intervention were carried out.

RESULTSAfter intervention, BMI ((25.53 ± 2.77) kg/m(2)), WHR (0.82 ± 0.03), TC ((4.60 ± 0.98) mmol/L), TG ((1.26 ± 0.88) mmol/L) in the intervention group were decreased significantly compared to the levels of BMI ((26.60 ± 3.18) kg/m(2)), WHR (0.93 ± 0.05), TC ((4.97 ± 1.02) mmol/L), TG ((1.98 ± 1.11) mmol/L) in the control group (all P values < 0.05); while HDL-C ((1.34 ± 0.26) mmol/L) in the intervention group was increased significantly compared to the level of HDL-C ((1.18 ± 0.17) mmol/L) in the control group (P < 0.05); After intervention, levels of T-AOC (19.52 ± 0.81), SOD ((85.42 ± 21.65) U/ml) and GSH-Px ((128.26 ± 33.65) µmol/L) were increased significantly compared to the levels of T-AOC (11.11 ± 1.30), SOD ((78.68 ± 30.48) U/ml) and GSH-Px ((118.48 ± 24.19) µmol/L) in the control group (all P values < 0.05); while MDA ((1.78 ± 1.16) nmol/ml) decreased significantly compared to the level of MDA ((2.12 ± 1.37) nmol/ml in the control group (P < 0.05); Pearson product moment correlation analysis showed that: T-AOC with TC, TG, BMI showed a negative correlation (r values were -0.258, -0.266, -0.230, respectively, all P values < 0.05), while with HDL-C was a positive correlation (r values was 0.194, P < 0.05); SOD with TC, TG, BMI showed a negative correlation (r values were -0.282, -0.311, -0.217, respectively, all P values < 0.05), while with HDL-C was a positive correlation (r values was 0.169, P < 0.05).

CONCLUSIONCompound whole grain could improve lipid metabolism to the hyperlipidemia population. There was a correlation between common human metabolism and the levels of oxidative stress.

Aged ; Aged, 80 and over ; Antioxidants ; China ; epidemiology ; Diet ; Diet Surveys ; Edible Grain ; Female ; Health Education ; Humans ; Hyperlipidemias ; epidemiology ; metabolism ; prevention & control ; Lipid Metabolism ; Male ; Middle Aged ; Oxidative Stress